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LAMPIRAN -2 HASIL PENGAMATAN
Larutan Standar Asam Galat
Polifenol total yang terukur merupakan total fenol yang terdapat pada ekstrak
teh putih berdasarkan standar asam galat (AG). Ekstraksi dilakukan dengan
cara reflux pada suhu 66oC menggunakan pelarut metanol 70 %. Larutan
standar yang digunakan yaitu 0.1 mg/g; 0,2 mg/gr; 0,4 mg/g; 0,8 mg/g; dan
1.6 mg/g dan diukur absorbansinya pada panjang gelombang 750 nm. kurva
baku dengan persamaan y =1.005 x - 0.001 (x konsestrasi AG (mg/g) dan y =
absorbansi) dan R² = 0.999.
1. Hasil Scan panjang gelombang optimum untuk Asam Galat Menggunakan Spektro fototemeter UV
stdr polyfenol as.galatAbs
Wavel ength (nm)400.0 600.0 800.0 1000.0
0.300
0.400
0.500
0.600
Position Height750 nm 0.643
2. Hasil Pengukuran Larutan Standar Asam GalatKonsentrasi (mg/gram)
(X) Absorbansi (Y)
0.1 0.0760.2 0.2290.4 0.3980.8 0.8011.6 1.608
750
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No. (X) (Y) XY X2 Y2
1 0.1 0.08 0.008 0.010 0.0062 0.2 0.23 0.046 0.040 0.0523 0.4 0.40 0.159 0.160 0.1584 0.8 0.80 0.641 0.640 0.6425 1.6 1.61 2.573 2.560 2.586∑ 3.1 3.11 3.426 3.410 3.444
y = a +b xa=(∑Y )¿¿
a=(3.112) (3.41 )−(3.112)(2.426)
9(3.41)−(3.1 ) ❑❑2
a = 0,001
b=n∑ XY −(∑ X ) (∑Y )
n∑ X2−¿¿
b=(3.426 )−(3.1 )(3.11)9(3.41)−(3.1 ) ❑❑
2
b = 1,005
r=n∑ XY−(∑ X ) (∑Y )
√¿¿¿
r=5(3,42)− (3,1 )(3,1)
√¿¿¿
r=7,4827,487
r = 0,999R2 = 0,9992
R2 = 0,999
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.80
0.20.40.60.8
11.21.41.61.8
f(x) = 1.00588709677419 x − 0.00124999999999931R² = 0.999067252621833
Kurva Standar Asam Galat
KonsentrasiLinear (Konsentrasi)
Konsentrasi (mg/gram)
Abso
rban
si
3. Analisis Pendahuluan
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Nama Uji : Uji Potensi PolifenolMetode : Follin Ciocalteu (Kulisic et al., 2006)Alat uji : Spektrofotomer UVReagen : Larutan Folin-Ciocalteau (1:1 (FeCl3 0,1 M; K3Fe(CN)6
0,008 M)Nama Sampel : Teh PutihKode Sampel : A, B, C, DPreparasi : Ekstraksi
Pengekstrak : Metanol 70 %Suhu : 66oCKondisi Ektrak : Refluks
Hasil Analisis Penelitian Pendahuluan Pendahuluan
No
Kode sampel C awal C awal
(mg/g)C alat (mg/g)
C alat rata2 Pengenceran
C alat rata2 total
Kadar Air
Kadar polyfenol
(%)1
A 0.500/101 g 4.95
0.55910.5670 2 1.1341 9.2342 25.03
2 0.57503
B 0.500/101 g 4.95
0.47210.46167 2 0.9233 9.8736 20.50
4 0.45125
C 0.500/101 g 4.95
0.47590.47295 2 0.9459 10.1863 21.06
6 0.47007
D 0.500/101 g 4.95
0.58680.5706 2 1.1411 9.6694 25.28
8 0.5543
4. Analisis Utama Polifenol TotalMetode : Follin CiocalteuNama Sampel : Teh PutihKode Sampel : Suhu (0C) Waktu (menit) KODE PENGUJIAN
553 t1w1
6 t1w2
9 t1w3
753 t2w1
6 t2w2
9 t2w2
953 t3w1
6 t3w2
9 t3w3
Preparasi : Diseduh (mengacu kepada SNI 01-1902-1995 )
Penyeduh : AquadesSuhu : 95oC (suhu didih),75, dan 85.Kondisi : Muka tempat seduh di tutup
Konsentrasi Larutan Standar = mg/gramKosentrasi Awal Seduhan Teh : 2,84g/140mL
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BJ air = 1 gram/mlBerat Teh = 2,84 gram (kadar air 9,67 %)
= 284 mgKosentrasi Awal = 284/140
= 20,89 mg/gKonsentrasi Kering = 18.33 mg/g
Pengenceran dilakukan karena pada konsentrasi 20,89 gram/gram melebihi absorbansi terukur melebihi larutan standar. sehingga diencerkan 2 kali.
Hasil Pengukuran SpektrofotometerNo Sampel Absorbansi (X) Konsentrasi Standar Eror Konsentrasi (Y)1 t1w1 0.121 0.1215 0.000 0.12152 t1w1 0.110 0.1106 0.000 0.11063 t1w2 0.224 0.2239 0.000 0.22394 t1w2 0.241 0.2408 0.000 0.24085 t1w3 0.233 0.2329 0.000 0.23296 t1w3 0.238 0.2379 0.000 0.23797 t2w1 0.314 0.2886 0.000 0.28868 t2w1 0.304 0.2836 0.000 0.28369 t2w2 0.287 0.3581 0.000 0.3581
10 t2w2 0.290 0.3552 0.000 0.355211 t2w3 0.449 0.4426 0.000 0.442612 t2w3 0.459 0.4476 0.000 0.447613 t3w1 0.258 0.2587 0.000 0.258714 t3w1 0.249 0.2577 0.000 0.257715 t3w2 0.423 0.4218 0.000 0.421816 t3w2 0.404 0.4029 0.000 0.402917 t3w3 0.549 0.5470 0.000 0.547018 t3w3 0.557 0.5550 0.000 0.5550
Contoh perhitungan Secara manualPersamaan Kurva Standar : y = 1.005x - 0.001Absorbansi t1w1 = 0.1215y = 1.005 (0.1215) – 0.001y = 0.1225 – 0.001 y = 0.1215
Konsentrasi polifenol total rata-rata t1w1 = 0,1162 mg/gKonsentrasi polifenol pada seduhan sebelum diencerkan = 0,1161 x 2
= 0,2321 mg/gKadar Polifenol dalam seduhan = (0,2321/(18,33)*(100)
= 1,44 %
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Hasil Uji Penelitian Utama
No Kode sampel
Konsentrasi AwalPengenceran
Konsentrasi Polifenol Terukur Spektro Konsentrasi Polifenol dalam
seduhan awalKadar polifenol pada
seduhan (%AG)Gram teh/ml air
Teh Kering (mg/g) (mg/g) Rata-rata (mg/g)
1 t1w1 2,84g/140mL 18.33 2 0.1215 0.1161 0.2321 1.142 0.11063 t1w2 2,84g/140mL 18.33 2 0.2239 0.2324 0.4647 2.294 0.24085 t1w3 2,84g/140mL 18.33 2 0.2329 0.2354 0.4708 2.326 0.23797 t2w1 2,84g/140mL 18.33 2 0.2886 0.2861 0.5722 2.828 0.28369 t2w2 2,84g/140mL 18.33 2 0.3581 0.3567 0.7133 3.5210 0.355211 t2w3 2,84g/140mL 18.33 2 0.4426 0.4451 0.8902 4.3912 0.447613 t3w1 2,84g/140mL 18.33 2 0.2587 0.2582 0.5164 2.5514 0.257715 t3w2 2,84g/140mL 18.33 2 0.4218 0.4124 0.8247 4.0616 0.402917 t3w3 2,84g/140mL 18.33 2 0.5470 0.5510 1.1020 5.4318 0.5550
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5. Hasil Scan Panjang Gelombang Optimum untuk DPPH kurva standar DPPHAbs
Wavel ength (nm)400.0 500.0 600.0 700.0 800.0 900.0
0.000
0.200
0.400
0.600
0.000
Position Height516.0 0.713
6. Penelitian Utama Penangkapan Radikal Bebas DPPHNama Uji : Uji Aktivitas AntioksidaMetode : DPPH (1,1-diphenyl- 2- Picrilhydrazl)Reagen : Larutan 1,1-Diphenyl-2-picrylhydrazyl (DPPH, Mr =
395,34) dengan konsentrasi akhir 2,0x10-4 M (Dibuat larutan stok pada konsentrasi 1,0x10-3 M)
Alat uji : Spektrofotomer UVNama Sampel : Teh PutihKode Sampel : Suhu (0C) Waktu (menit) KODE PENGUJIAN
55 3 t1w1
55 6 t1w2
55 9 t1w3
75 3 t2w1
75 6 t2w2
75 9 t2w2
95 3 t3w1
95 6 t3w2
95 9 t3w3
Preparasi : DiseduhPenyeduh : AquadesSuhu : 95oC (suhu didih),75, dan 85.Kondisi : Muka tempat seduh di tutup
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Hasil Uji DPPH1. Kode t1w1
Tabel Absorbansi Kode t1w1 Absorbansi rata-rata
C sampel (ppm)
% Inhibisi
0.8550.276 162.2857 67.720.558 81.1429 34.740.705 40.5714 17.540.799 20.2857 6.550.828 5.0715 3.16
0 20 40 60 80 100 120 140 160 1800.00
10.0020.0030.0040.0050.0060.0070.0080.00
f(x) = 0.418816575650471 x + 0.02873583647272R² = 0.997924994122672
Kurva daya hambat sampel T1W1 terhadap radikal bebas DPPH
C sampel (ppm)
% in
hibi
si
Contoh Perhitungan Inhibisi
% Inhibisi =A referece−A sample
A referenceA reference = 0,855 A Sampel = 0,267
% Inhibisi = 0.855−0.267
0.855 x 100
= 67.72 EC50 = Persen Inhibisi 50%y = 50y = 0,418x + 0,02850 = 0,418x + 0,0280,418x = 50 - 0,028x = 49,972/0,418
= 119.5502 ppm
2. Kode t1w2
Tabel Absorbansi Kode t1w2
Absorbansi rata-rata
C sampel (ppm)
% Inhibisi
0.862 0.429 81.1429 50.230.661 40.5714 23.320.761 20.2857 11.720.826 10.1429 4.18 0 10 20 30 40 50 60 70 80 90
0.00
10.00
20.00
30.00
40.00
50.00
60.00
f(x) = 0.642391444643165 x − 2.07303546769097R² = 0.999075040971165
Kurva daya hambat sampel T1W2 terhadap radikal bebas DPPH
C sampel (ppm)
% In
hibi
si
EC50 = Persen Inhibisi 50%y = 50y = 0,642x - 2,07350 = 0,642x - 2,0730,642 x = 50 + 2,073x = 52,073 / 0,642
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= 88,111 ppm3. Kode t1w3
Tabel Absorbansi Kode t1w3
Absorbansi rata-rata
C sampel (ppm) % Inhibisi
0.862 0.418 81.1429 51.510.616 40.5714 28.540.769 20.2857 10.790.812 10.1429 5.80 0 10 20 30 40 50 60 70 80 90
0.00
10.00
20.00
30.00
40.00
50.00
60.00
f(x) = 0.655718621528408 x − 0.781800451216405R² = 0.991252689881529
Kurva daya hambat sampel T1W3 terhadap radikal bebas DPPH
C sampel (ppm)
% In
hibi
si
EC50 = Persen Inhibisi 50%y = 50y = 0,655x – 0.78150 = 0,655x – 0.7810.655x = 50+0,781x = 50,781 /0,655
= 88,111 ppmEc50 T1w3 = 88,111 ppm
4. Kode t2w1
Tabel Absorbansi Kode t2w1 Absorbansi rata-rata
C sampel (ppm)
% Inhibisi
0.822 0.353 81.1429 57.060.559 40.5714 32.000.723 20.2857 12.040.749 10.1429 8.88
0 10 20 30 40 50 60 70 80 900.00
10.00
20.00
30.00
40.00
50.00
60.00f(x) = 0.702958687497473 x + 0.75637395092436R² = 0.987991214759008
Kurva daya hambat sampel T2W1 terhadap radikal bebas DPPH
C sampel (ppm)
% In
hibi
si
EC50 = Persen Inhibisi 50% y = 50y = 0,703x + 0,76550 = 0,703x + 0,7650.703x = 50-0,765x = 49,235/0,703
= 70.048 ppmEc50 T2w1 = 70.048 ppm
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5. Kode T2w2
Tabel Absorbansi Kode t2w2
Absorbansi rata-rata
C sampel (ppm)
% Inhibisi
0.8280.179 81.1429 78.380.469 40.5714 43.360.673 20.2857 18.720.745 10.1429 10.020.792 5.0715 4.35
0 10 20 30 40 50 60 70 80 900.00
20.00
40.00
60.00
80.00
100.00
f(x) = 0.980999121556703 x + 0.12074591566639R² = 0.995712513507543
Kurva daya hambat sampel T2W2 terhadap radikal bebas DPPH
C sampel (ppm)
% in
hibi
si
EC50 = Persen Inhibisi 50% y = 50y = 0,981x + 0,12050 = 0,981x + 0,1200.981x = 50-0,120x = 49,880 /0,981
= 49.877 ppmEc50 T2w2 = 49.877 ppm
6. Kode t2w3
Tabel Absorbansi Kode t2w3
Absorbansi rata-rata
C sampel (ppm)
% Inhibisi
0.823 0.129 81.1429 84.330.446 40.5714 45.810.637 20.2857 22.600.727 10.1429 11.660.784 5.0715 4.74
0 5 10 15 20 25 30 35 40 450.00
10.00
20.00
30.00
40.00
50.00
f(x) = 1.14462135427851 x − 0.565349677610506R² = 0.999337027297215
Kurva daya hambat sampel T2W3 terhadap radikal bebas DPPH
C sampel (ppm)
% In
hibi
si
EC50 = Persen Inhibisi 50% y = 50y = 1,144x - 0,56550 = 1,144x - 0,5651.144x = 50 + 0,565x = 50,565 /1,144
= 44,200 ppmEc50 T2w3 = 44,200 ppm
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7. Kode t3w1
Tabel Absorbansi Kode t3w1
Absorbansi rata-rata
C sampel (ppm)
% Inhibis
i0.844 0.407 162.2857 51.780.398 81.1429 52.840.624 40.5714 26.070.753 20.2857 10.78
0.800 10.1429 5.210 10 20 30 40 50 60 70 80 90
0.00
10.00
20.00
30.00
40.00
50.00
60.00
f(x) = 0.678642033412916 x − 2.08633843879901R² = 0.998986065263011
Kurva daya hambat sampel T3w1 terhadap radikal bebas DPPH
C sampel (ppm)
% In
hibi
siEC50 = Persen Inhibisi 50% y = 50y =0,678x - 2,08650 =0,678x - 2,0860,678x = 50 + 2,086x = 52,086 /1,144
= 76,755 ppmEc50 T3w1 = 76,755 ppm
8. Kode t3w2
Tabel Absorbansi Kode t3w2
Absorbansi rata-rata
C sampel (ppm)
% Inhibisi
0.840 0.163 81.1429 80.600.476 40.5714 43.330.669 20.2857 20.360.754 10.1429 10.24 0 10 20 30 40 50 60 70 80 90
0.0010.0020.0030.0040.0050.0060.0070.0080.0090.00
f(x) = 0.993059488138045 x + 0.859214781492959R² = 0.997816530949792
Kurva daya hambat sampel T3w2 terhadap radikal bebas DPPH
C sampel (ppm)
% In
hibi
si
EC50 = Persen Inhibisi 50% y = 50y =0,993x + 0,85950 =0,993x + 0,8590,993x = 50 - 0,859x = 49,141 /0,993
= 49,482 ppmEc50 T3w2 = 49,482 ppm
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9. Kode t3w3
Tabel Absorbansi Kode t2w3
Absorbansi rata-rata
C sampel (ppm)
% Inhibisi
0.838 0.074 81.1429 91.170.365 40.5714 56.440.581 20.2857 30.670.681 10.1429 18.740.752 5.0715 10.26
0 5 10 15 20 25 30 35 40 450.00
10.0020.0030.0040.0050.0060.00
f(x) = 1.27943265234612 x + 4.69535604747913R² = 0.998347051130522
Kurva daya hambat sampel T3w3 terhadap radikal bebas DPPH
C sampel (ppm)
% In
hibi
si
EC50 = Persen Inhibisi 50% y = 50y =1,279x + 4,69550 =1,279x + 4,6951,279x = 50 - 4,695x = 49,141 /1,279
= 35,411 ppmEc50 T3w1 = 35,411 ppm
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LAMPIRAN -3 ANALISIS DATA
1. Analisis Polifenol TotalNo. X1 X2 Y
1 55 3 1.272 55 6 2.543 55 9 2.574 75 3 3.125 75 6 3.896 75 9 4.867 95 3 2.828 95 6 4.59 95 9 6.01
Keterangan :X1 = Suhu penyeduhan (oC)X2 = Lama Penyeduhan (menit)Y = % Polifenol Total
Statistik Deskrivtif
Mean Std. Deviation N
Polifenol_Total 3.5089 1.44340 9
suhu 75.0000 17.32051 9
waktu 6.0000 2.59808 9
Korelasi Parsial
Polifenol_Total suhu waktu
Pearson Correlation Polifenol_Total 1.000 .695 .623
suhu .695 1.000 .000
waktu .623 .000 1.000
Sig. (1-tailed) Polifenol_Total . .019 .037
suhu .019 . .500
waktu .037 .500 .
N Polifenol_Total 9 9 9
suhu 9 9 9
waktu 9 9 9
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Variables Entered/Removedb
Model
Variables
Entered
Variables
Removed Method
1 waktu, suhua . Enter
a. All requested variables entered.
b. Dependent Variable: Polifenol_Total
Model Summaryb
Model R R Square
Adjusted R
Square
Std. Error of the
Estimate
1 .933a .871 .828 .59834
a. Predictors: (Constant), waktu, suhu
b. Dependent Variable: Polifenol_Total
ANOVAb
Model Sum of Squares df Mean Square F Sig.
1 Regression 14.519 2 7.260 20.278 .002a
Residual 2.148 6 .358
Total 16.667 8
a. Predictors: (Constant), waktu, suhu
b. Dependent Variable: Polifenol_Total
Coefficientsa
Model
Unstandardized Coefficients
Standardized
Coefficients
t Sig.B Std. Error Beta
1 (Constant) -2.912 1.057 -2.754 .033
suhu .058 .012 .695 4.742 .003
waktu .346 .081 .623 4.251 .005
a. Dependent Variable: Polifenol_Total
69
2. Analisis Penangkapan DPPHNo. X1 X2 Y
1 55 3 119.552 55 6 81.113 55 9 77.534 75 3 70.055 75 6 49.886 75 9 44.27 95 3 76.768 95 6 49.489 95 9 35.41
Keterangan :X1 = Suhu penyeduhan (oC)X2 = Lama Penyeduhan (menit)Y = EC50 DPPH
Descriptive Statistics
Mean Std. Deviation N
DPPH 67.1078 25.72388 9
suhu 80.0000 17.32051 9
waktu 6.0000 2.59808 9
Correlations
DPPH suhu waktu
Pearson Correlation DPPH 1.000 -.654 -.613
suhu -.654 1.000 .000
waktu -.613 .000 1.000Sig. (1-tailed) DPPH . .028 .040
suhu .028 . .500waktu .040 .500 .
N DPPH 9 9 9
suhu 9 9 9
waktu 9 9 9
Variables Entered/Removedb
Model
Variables
Entered
Variables
Removed Method
1 waktu, suhua . Enter
a. All requested variables entered.
b. Dependent Variable: DPPH
70
Model Summaryb
Model R R Square
Adjusted R
Square
Std. Error of the
Estimate
1 .896a .803 .738 13.17814
a. Predictors: (Constant), waktu, suhu
b. Dependent Variable: DPPH
ANOVAb
Model Sum of Squares df Mean Square F Sig.
1 Regression 4251.763 2 2125.882 12.241 .008a
Residual 1041.981 6 173.663
Total 5293.744 8
a. Predictors: (Constant), waktu, suhu
b. Dependent Variable: DPPH
Coefficientsa
Model
Unstandardized Coefficients
Standardized
Coefficients
t Sig.B Std. Error Beta
1 (Constant) 181.208 24.458 7.409 .000
suhu -.971 .269 -.654 -3.610 .011
waktu -6.068 1.793 -.613 -3.384 .015
a. Dependent Variable: DPPH
71
3. Korelasi Plifenol Total dan Penengkapan Radikal Bebas DPPHNo. Y X
1 119.55 1.272 81.11 2.543 77.53 2.574 70.05 3.125 49.88 3.896 44.2 4.867 76.76 2.828 49.48 4.59 35.41 6.01
Keterangan :
Y = EC50 DPPH
X = Polifenol total Descriptive Statistics
Mean Std. Deviation N
DPPH 67.1078 25.72388 9
Polifenol_Total 3.5089 1.44340 9
Correlations
DPPH Polifenol_Total
Pearson Correlation DPPH 1.000 -.952
Polifenol_Total -.952 1.000Sig. (1-tailed) DPPH . .000
Polifenol_Total .000 .N DPPH 9 9
Polifenol_Total 9 9
Variables Entered/Removedb
Model
Variables
Entered
Variables
Removed Method
1 Polifenol_Totala . Enter
a. All requested variables entered.
b. Dependent Variable: DPPH
72
Model Summaryb
Model R R Square
Adjusted R
Square
Std. Error of the
Estimate
1 .952a .907 .893 8.40510
a. Predictors: (Constant), Polifenol_Total
b. Dependent Variable: DPPH
ANOVAb
Model Sum of Squares df Mean Square F Sig.
1 Regression 4799.224 1 4799.224 67.934 .000a
Residual 494.520 7 70.646
Total 5293.744 8
a. Predictors: (Constant), Polifenol_Total
b. Dependent Variable: DPPH
Coefficientsa
Model
Unstandardized Coefficients
Standardized
Coefficients
t Sig.B Std. Error Beta
1 (Constant) 126.650 7.748 16.345 .000
Polifenol_Total -16.969 2.059 -.952 -8.242 .000
a. Dependent Variable: DPPH